Radial Flow Immobilized Metal Ion Affinity Monolithic Column And Its Chromatographic Performance

Monolithic column was prepared by in situ polymerization inside the tube, andthen, according to application, derivatived the monolithic polymer materials to getthe monolithic column for the application of separation analysis, enrichment orpreparation. Compared to the conventional packed column, monolithic column hadsome merits, simple preparation method, good permeability, low pre-columnpressure, high utilization of the active site, high column capacity, etc, and itsresolution and the adsorption capacity was not dependent on flow rate, when highvelocity operated the pressure drop was low, especially favorable to the preparationand separation for biological samples. Therefore, to study preparation,characterization and application of monolith column have important theoretical andapplication value.Monolithic supports are considered as the fourth generation of sorbent materialsfor protein separations,and their internal structure with highly interconnected poresconsists of macropores (pore diameters﹥50nm), mesopores (pore diametersbetween50nm and2nm) and micropores (pore diameters﹤2nm).The macroporescan achieve convective mass transfer between the mobile and the stationary phases,which substantially increase the mobility of the molecules to be separated comparedto traditional bead counterparts. While mesopores and micropores can afford activesites and larger surface of monolithic supports at the same time，increase columncapacity and improve chromatographic separation capabilities.In this paper, we reviewed the history of the development, preparation processand application of the monolithic column at first. Study the preparation method ofmonolithic column, and prepared the metal chelation chromatography monolithiccolumn, characterized the monolithic column by the SEM, IR, EDS, for theproportion of monomer, crosslinker and porogenic agent, combined the way tocharacterized, optimized the preparation conditions of the monolithic column. Observed by SEM, the morphology of the different positions of the monolithicmatrix is more similar, showing the throughpore network。 IR characterizationshowed that the surface have been modified by IDA and EDS showed that thematerials was already chelation. To research the chromatographic properties ofmonolithic columns, we changed the chromatographic conditions such as pH value,gradient, flow rate and other parameters, to investigate the influence of theseparameters on the monolithic stationary phases resolution. The results show that thepH value, flow rate did not affect on the resolution of the monolithic column and theretention time, to investigate the reproducibility of different batches of thepreparation of monolithic column, we determinated the monolithic stationary phasesdynamic column capacity and and a flow rate of the column capacity by Head-onanalysis method. All the results proved the superiority of the monolithic material,short analysis time, big column capacity, reflecting the convenience of theseparation of biological macromolecules.